Interactions of Influenza Virus with Cultured Cells: Detailed Kinetic Modeling of Binding and Endocytosis

Detalhes bibliográficos
Autor(a) principal: Nunes-Correia, Isabel
Data de Publicação: 1999
Outros Autores: Ramalho-Santos, João, Nir, Shlomo, Lima, Maria C. Pedroso de
Tipo de documento: Artigo
Idioma: eng
Título da fonte: Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos)
Texto Completo: http://hdl.handle.net/10316/10556
https://doi.org/10.1021/bi9812524
Resumo: We performed a detailed kinetic analysis of the uptake of influenza virus (A/PR8/34) by Madin Darby canine kidney (MDCK) cells in culture. Experimental procedures were based on the relief of fluorescence self-quenching of the fluorescent probe octadecylrhodamine B chloride (R18) incorporated in the viral envelope. Equilibrium for binding of influenza virus to MDCK cells (2.5 × 106/mL) was reached quicker with temperature increases due to a faster dynamic mobility of the particles. We deduced that there are two kinds of binding sites for influenza virus in MDCK cells and determined the kinetic parameters of the binding process (adhesion and detachment rate constants), using a mass action kinetic model. As the temperature increases, the number of binding sites for influenza virus decreases, especially the high-affinity binding sites, whereas the value of the affinity constant for virus binding to the binding site, k, increases. Nevertheless, the binding association constant at equilibrium Ki, which is given by Ki = Niki, where Ni is the number of binding sites per cell, declines as the temperature increases. When endocytosis occurs, the total uptake of virions by the cells is larger than that observed in the process of binding at the same temperature, and the uptake proceeds for longer times. Using our mass kinetic model, we determined that at 20 °C, the rate constant of endocytosis, ε, for influenza virus with this cell line is 2.6 × 10-4 s-1, i.e., in the same range as in studies on endocytosis of liposomes.
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spelling Interactions of Influenza Virus with Cultured Cells: Detailed Kinetic Modeling of Binding and EndocytosisWe performed a detailed kinetic analysis of the uptake of influenza virus (A/PR8/34) by Madin Darby canine kidney (MDCK) cells in culture. Experimental procedures were based on the relief of fluorescence self-quenching of the fluorescent probe octadecylrhodamine B chloride (R18) incorporated in the viral envelope. Equilibrium for binding of influenza virus to MDCK cells (2.5 × 106/mL) was reached quicker with temperature increases due to a faster dynamic mobility of the particles. We deduced that there are two kinds of binding sites for influenza virus in MDCK cells and determined the kinetic parameters of the binding process (adhesion and detachment rate constants), using a mass action kinetic model. As the temperature increases, the number of binding sites for influenza virus decreases, especially the high-affinity binding sites, whereas the value of the affinity constant for virus binding to the binding site, k, increases. Nevertheless, the binding association constant at equilibrium Ki, which is given by Ki = Niki, where Ni is the number of binding sites per cell, declines as the temperature increases. When endocytosis occurs, the total uptake of virions by the cells is larger than that observed in the process of binding at the same temperature, and the uptake proceeds for longer times. Using our mass kinetic model, we determined that at 20 °C, the rate constant of endocytosis, ε, for influenza virus with this cell line is 2.6 × 10-4 s-1, i.e., in the same range as in studies on endocytosis of liposomes.American Chemical Society1999-01-19info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/articlehttp://hdl.handle.net/10316/10556http://hdl.handle.net/10316/10556https://doi.org/10.1021/bi9812524engBiochemistry. 38:3 (1999) 1095-11010006-2960Nunes-Correia, IsabelRamalho-Santos, JoãoNir, ShlomoLima, Maria C. Pedroso deinfo:eu-repo/semantics/openAccessreponame:Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos)instname:Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informaçãoinstacron:RCAAP2021-09-17T11:05:01Zoai:estudogeral.uc.pt:10316/10556Portal AgregadorONGhttps://www.rcaap.pt/oai/openaireopendoar:71602024-03-19T20:55:46.919656Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) - Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informaçãofalse
dc.title.none.fl_str_mv Interactions of Influenza Virus with Cultured Cells: Detailed Kinetic Modeling of Binding and Endocytosis
title Interactions of Influenza Virus with Cultured Cells: Detailed Kinetic Modeling of Binding and Endocytosis
spellingShingle Interactions of Influenza Virus with Cultured Cells: Detailed Kinetic Modeling of Binding and Endocytosis
Nunes-Correia, Isabel
title_short Interactions of Influenza Virus with Cultured Cells: Detailed Kinetic Modeling of Binding and Endocytosis
title_full Interactions of Influenza Virus with Cultured Cells: Detailed Kinetic Modeling of Binding and Endocytosis
title_fullStr Interactions of Influenza Virus with Cultured Cells: Detailed Kinetic Modeling of Binding and Endocytosis
title_full_unstemmed Interactions of Influenza Virus with Cultured Cells: Detailed Kinetic Modeling of Binding and Endocytosis
title_sort Interactions of Influenza Virus with Cultured Cells: Detailed Kinetic Modeling of Binding and Endocytosis
author Nunes-Correia, Isabel
author_facet Nunes-Correia, Isabel
Ramalho-Santos, João
Nir, Shlomo
Lima, Maria C. Pedroso de
author_role author
author2 Ramalho-Santos, João
Nir, Shlomo
Lima, Maria C. Pedroso de
author2_role author
author
author
dc.contributor.author.fl_str_mv Nunes-Correia, Isabel
Ramalho-Santos, João
Nir, Shlomo
Lima, Maria C. Pedroso de
description We performed a detailed kinetic analysis of the uptake of influenza virus (A/PR8/34) by Madin Darby canine kidney (MDCK) cells in culture. Experimental procedures were based on the relief of fluorescence self-quenching of the fluorescent probe octadecylrhodamine B chloride (R18) incorporated in the viral envelope. Equilibrium for binding of influenza virus to MDCK cells (2.5 × 106/mL) was reached quicker with temperature increases due to a faster dynamic mobility of the particles. We deduced that there are two kinds of binding sites for influenza virus in MDCK cells and determined the kinetic parameters of the binding process (adhesion and detachment rate constants), using a mass action kinetic model. As the temperature increases, the number of binding sites for influenza virus decreases, especially the high-affinity binding sites, whereas the value of the affinity constant for virus binding to the binding site, k, increases. Nevertheless, the binding association constant at equilibrium Ki, which is given by Ki = Niki, where Ni is the number of binding sites per cell, declines as the temperature increases. When endocytosis occurs, the total uptake of virions by the cells is larger than that observed in the process of binding at the same temperature, and the uptake proceeds for longer times. Using our mass kinetic model, we determined that at 20 °C, the rate constant of endocytosis, ε, for influenza virus with this cell line is 2.6 × 10-4 s-1, i.e., in the same range as in studies on endocytosis of liposomes.
publishDate 1999
dc.date.none.fl_str_mv 1999-01-19
dc.type.status.fl_str_mv info:eu-repo/semantics/publishedVersion
dc.type.driver.fl_str_mv info:eu-repo/semantics/article
format article
status_str publishedVersion
dc.identifier.uri.fl_str_mv http://hdl.handle.net/10316/10556
http://hdl.handle.net/10316/10556
https://doi.org/10.1021/bi9812524
url http://hdl.handle.net/10316/10556
https://doi.org/10.1021/bi9812524
dc.language.iso.fl_str_mv eng
language eng
dc.relation.none.fl_str_mv Biochemistry. 38:3 (1999) 1095-1101
0006-2960
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dc.publisher.none.fl_str_mv American Chemical Society
publisher.none.fl_str_mv American Chemical Society
dc.source.none.fl_str_mv reponame:Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos)
instname:Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informação
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